CN1337401A - Thiophosphoryl amino acid ester compound and its prepn process - Google Patents
Thiophosphoryl amino acid ester compound and its prepn process Download PDFInfo
- Publication number
- CN1337401A CN1337401A CN 01130784 CN01130784A CN1337401A CN 1337401 A CN1337401 A CN 1337401A CN 01130784 CN01130784 CN 01130784 CN 01130784 A CN01130784 A CN 01130784A CN 1337401 A CN1337401 A CN 1337401A
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- Prior art keywords
- isopropylidene
- compound
- nmr
- solution
- 1mmol
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Links
- -1 Thiophosphoryl amino acid ester compound Chemical class 0.000 title claims abstract description 50
- 238000000034 method Methods 0.000 title claims description 3
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims abstract description 142
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims abstract description 71
- 238000006243 chemical reaction Methods 0.000 claims abstract description 68
- 150000001875 compounds Chemical class 0.000 claims abstract description 65
- 125000003253 isopropoxy group Chemical group [H]C([H])([H])C([H])(O*)C([H])([H])[H] 0.000 claims abstract description 48
- 238000002360 preparation method Methods 0.000 claims abstract description 37
- 238000004440 column chromatography Methods 0.000 claims abstract description 23
- 238000000926 separation method Methods 0.000 claims abstract description 23
- 239000002777 nucleoside Substances 0.000 claims abstract description 20
- 239000002994 raw material Substances 0.000 claims abstract description 13
- 239000002253 acid Substances 0.000 claims abstract description 8
- 239000011230 binding agent Substances 0.000 claims abstract description 7
- 238000003756 stirring Methods 0.000 claims description 52
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 44
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 42
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 238000004821 distillation Methods 0.000 claims description 23
- 238000009835 boiling Methods 0.000 claims description 22
- 239000000463 material Substances 0.000 claims description 22
- 238000012544 monitoring process Methods 0.000 claims description 22
- 229910052757 nitrogen Inorganic materials 0.000 claims description 22
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 10
- 229940104302 cytosine Drugs 0.000 claims description 7
- 239000011782 vitamin Substances 0.000 claims description 7
- 235000013343 vitamin Nutrition 0.000 claims description 7
- 229940088594 vitamin Drugs 0.000 claims description 7
- 229930003231 vitamin Natural products 0.000 claims description 7
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 7
- 238000005352 clarification Methods 0.000 claims description 6
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 claims description 6
- GKASDNZWUGIAMG-UHFFFAOYSA-N triethyl orthoformate Chemical compound CCOC(OCC)OCC GKASDNZWUGIAMG-UHFFFAOYSA-N 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 4
- 125000003835 nucleoside group Chemical group 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 230000006837 decompression Effects 0.000 claims description 3
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 239000003292 glue Substances 0.000 claims description 2
- 150000004702 methyl esters Chemical class 0.000 claims description 2
- 229910052710 silicon Inorganic materials 0.000 claims description 2
- 239000010703 silicon Substances 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 abstract description 4
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 126
- 238000005481 NMR spectroscopy Methods 0.000 description 60
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 40
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 30
- 239000000047 product Substances 0.000 description 28
- 238000002474 experimental method Methods 0.000 description 25
- 230000000694 effects Effects 0.000 description 22
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 20
- 238000005160 1H NMR spectroscopy Methods 0.000 description 20
- 238000004679 31P NMR spectroscopy Methods 0.000 description 20
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 20
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 20
- 230000015572 biosynthetic process Effects 0.000 description 20
- 239000003480 eluent Substances 0.000 description 20
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 20
- 229910002027 silica gel Inorganic materials 0.000 description 20
- 239000000741 silica gel Substances 0.000 description 20
- 229960001866 silicon dioxide Drugs 0.000 description 20
- 230000003595 spectral effect Effects 0.000 description 20
- 238000003786 synthesis reaction Methods 0.000 description 20
- 230000007062 hydrolysis Effects 0.000 description 19
- 238000006460 hydrolysis reaction Methods 0.000 description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 18
- UBGDNZQSYVIVHU-PEBGCTIMSA-N 1-[(3ar,4r,6r,6ar)-6-(hydroxymethyl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]-4-aminopyrimidin-2-one Chemical compound N1([C@@H]2O[C@H](CO)[C@H]3OC(O[C@H]32)(C)C)C=CC(N)=NC1=O UBGDNZQSYVIVHU-PEBGCTIMSA-N 0.000 description 17
- XKPDAYWPKILAMO-UHFFFAOYSA-N 2',3'-O-Isopropylidene-Guanosine Natural products C1=NC(C(N=C(N)N2)=O)=C2N1C1OC(CO)C2OC(C)(C)OC21 XKPDAYWPKILAMO-UHFFFAOYSA-N 0.000 description 17
- XKPDAYWPKILAMO-IOSLPCCCSA-N 9-[(3ar,4r,6r,6ar)-6-(hydroxymethyl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]-2-amino-3h-purin-6-one Chemical compound C1=NC(C(N=C(N)N2)=O)=C2N1[C@@H]1O[C@H](CO)[C@H]2OC(C)(C)O[C@H]21 XKPDAYWPKILAMO-IOSLPCCCSA-N 0.000 description 17
- GFDUSNQQMOENLR-PEBGCTIMSA-N 1-[(3ar,4r,6r,6ar)-6-(hydroxymethyl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-4-yl]pyrimidine-2,4-dione Chemical compound N1([C@@H]2O[C@H](CO)[C@H]3OC(O[C@H]32)(C)C)C=CC(=O)NC1=O GFDUSNQQMOENLR-PEBGCTIMSA-N 0.000 description 16
- GFDUSNQQMOENLR-UHFFFAOYSA-N 2',3'-O-Isopropylidene-Uridine Natural products C12OC(C)(C)OC2C(CO)OC1N1C=CC(=O)NC1=O GFDUSNQQMOENLR-UHFFFAOYSA-N 0.000 description 16
- LCCLUOXEZAHUNS-WOUKDFQISA-N [(3ar,4r,6r,6ar)-4-(6-aminopurin-9-yl)-2,2-dimethyl-3a,4,6,6a-tetrahydrofuro[3,4-d][1,3]dioxol-6-yl]methanol Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]1O[C@H](CO)[C@H]2OC(C)(C)O[C@H]21 LCCLUOXEZAHUNS-WOUKDFQISA-N 0.000 description 15
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 15
- 235000001014 amino acid Nutrition 0.000 description 9
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 8
- 241000700605 Viruses Species 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 4
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 4
- 230000000840 anti-viral effect Effects 0.000 description 4
- COQRGFWWJBEXRC-UHFFFAOYSA-N hydron;methyl 2-aminoacetate;chloride Chemical compound Cl.COC(=O)CN COQRGFWWJBEXRC-UHFFFAOYSA-N 0.000 description 4
- DWKPPFQULDPWHX-VKHMYHEASA-N l-alanyl ester Chemical compound COC(=O)[C@H](C)N DWKPPFQULDPWHX-VKHMYHEASA-N 0.000 description 4
- KUGLDBMQKZTXPW-JEDNCBNOSA-N methyl (2s)-2-amino-3-methylbutanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)C(C)C KUGLDBMQKZTXPW-JEDNCBNOSA-N 0.000 description 4
- SWVMLNPDTIFDDY-FVGYRXGTSA-N methyl (2s)-2-amino-3-phenylpropanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)CC1=CC=CC=C1 SWVMLNPDTIFDDY-FVGYRXGTSA-N 0.000 description 4
- QVDXUKJJGUSGLS-LURJTMIESA-N methyl L-leucinate Chemical compound COC(=O)[C@@H](N)CC(C)C QVDXUKJJGUSGLS-LURJTMIESA-N 0.000 description 4
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical compound OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000005457 ice water Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 2
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 2
- 101710163270 Nuclease Proteins 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 229940029575 guanosine Drugs 0.000 description 2
- 150000004713 phosphodiesters Chemical class 0.000 description 2
- 229940002612 prodrug Drugs 0.000 description 2
- 239000000651 prodrug Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 2
- 229940045145 uridine Drugs 0.000 description 2
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 241000725303 Human immunodeficiency virus Species 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- QCJQWJKKTGJDCM-UHFFFAOYSA-N [P].[S] Chemical compound [P].[S] QCJQWJKKTGJDCM-UHFFFAOYSA-N 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 239000000074 antisense oligonucleotide Substances 0.000 description 1
- 238000012230 antisense oligonucleotides Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 125000003963 dichloro group Chemical group Cl* 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 125000004437 phosphorous atom Chemical group 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003582 thiophosphoric acids Chemical class 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Saccharide Compounds (AREA)
Abstract
The present invention relates to a thiophosphoryl amino acid ester compound. Said invention provides its structural formula, and its preparation method incldues the following steps: dissolving raw material (isopropoxy substituted) dichlorthion in tetrahydrofuran, adding amino acid methyl ester hydrochlorate to said solution, drop-adding acid-binding agent, and adding 2',3'-O-isopropylidene nucleoside solution in said solution, then more drop-adding acid-binding to make reaction, after reaction filtering, rotating distrillation, column chromatography to make separation, so that the invented compound can be obtained.
Description
The present invention relates to a kind of thiophosphoryl amino acid ester compound and preparation method thereof, relate in particular to 2 ', 3 '-O-isopropylidene nucleosides 5 '-preparation method of (isopropoxy replacement) thiophosphoryl amino acid ester compound, this compounds has good biological activity and antiviral, antitumor, anti-ly likes now pharmaceutical activity such as virus, can develop in clinical application that a kind of ucleosides is antiviral, antitumor, anti-likes viral now prodrug (prodrugs).
Thiophosphoric acid contains the structure that a sulphur replaces oxygen on phosphorus atom, though this structure is still keeping original electric charge, the character that is keeping highly water-soluble, but other physicochemical properties of the dna oligo of thiophosphoric acid (S-oligos) and biology attribute all have very big difference with natural phosphodiester prototype.One of marked difference is that thiophosphoric acid has resistivity to nuclease.Because the phosphodiester class antisense oligonucleotide that adds has been fallen in the existence of exonuclease in the cell, very fast digestion, make its forfeiture action function.By synthetic thiophosphoric acid compounds with opposing nuclease, be used for the treatment of purpose for using the synthetic dna oligo from now on, opened up the road of a hope beyond doubt.
The objective of the invention is to propose a kind of thiophosphoryl amino acid ester compound and preparation method thereof, specifically, be 2 ', 3 '-O-isopropylidene nucleosides 5 '-preparation method of (isopropoxy replacement) thiophosphoryl amino acid ester compound, to be used to develop antiviral, antitumor, the anti-active medicine of virus now of liking.
2 of the present invention's proposition ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound, its structural formula is:
R is H in the said structure formula, CH
3, C
6H
5CH
2, (CH
3)
2CH
2, (CH
3)
2CHCH
2B is a VITAMIN B4, guanine, cytosine(Cyt), uridylic.
The preparation method of above-claimed cpd comprises the steps:
1) under nitrogen protection, cryosel is bathed under the condition that is cooled to-4～-8 ℃, and raw material (isopropoxy replacement) two compd 22/190s are dissolved in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution that concentration is 0.8～1.0mol/L.
2) add amino acid methyl ester hydrochloride with the amount of raw material same substance in above-mentioned solution, slowly drip the acid binding agent triethylamine after stirring, the add-on of acid binding agent is 2 times of amount of raw material.
3) preparation 2 ', 3 '-O-isopropylidene nucleosides solution for standby:
Get nucleosides under the room temperature and be suspended in the solution that is made into 0.5～1.0mol/L in the exsiccant solvent, while stirring to wherein adding and the tosic acid of amount of substances such as nucleosides and the triethyl orthoformate of four times of amount of substances, after system becomes clarification, neutralize with big water gaging with strong aqua and to make PH=7～8, then above-mentioned solution decompression is concentrated, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene nucleosides.With make 2 ', 3 '-be dissolved in the solution that is mixed with 0.5～1.0mol/L in the solvent after the O-isopropylidene nucleosides suction filtration drying.
4) follow the tracks of above-mentioned second reaction process that goes on foot, after treating that the raw material total overall reaction finishes, will with 2 of above-mentioned the 3rd step preparation of the amount of raw material same substance ', 3 '-O-isopropylidene nucleosides solution slowly splashes in the solution in second step, and the back that stirs continue to drip the acid binding agent triethylamine with the amount of raw material same substance.
5) after monitoring reaction is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.Recycle silicon glue post carries out column chromatography for separation, can obtain 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound.
With 2 of method of the present invention preparation ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid methyl ester, be a class brand-new have an anti-active compound of virus drugs now of liking, by synthetic different types of nucleosides-amino acid conjugate, and carry out activity experiment, tentatively obtained satisfied result, liked now the virus drugs achievement in research that provides the foundation for further development and development of new resist.
Below introduce embodiments of the invention:
Embodiment 1: preparation 2 ', '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene adenosine is standby:
Getting 1mol (267g) adenosine under the room temperature is suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system becomes clarification after about 1 hour, neutralize with 100ml strong aqua and 5.9L water, then above-mentioned solution decompression is concentrated into 1.5L, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene adenosine 266g (87%).
4) follow the tracks of reaction process with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) after the reaction of monitoring above-mentioned second step with NMR is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.Carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 58.4%.
Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 60.38,61.02;
1H NMR (500MHz, DMSO-d
6): δ 9.26,9.22 (bs, 1H, NH), 8.43 (1H, s, H-2), 8.23 (1H, s, H-8), 7.40 (2H, s, NH
2), 6.44 (1H, m, H-1 '), 4,50 (2H, m, H-2 ', 3 '), 3.97 (1H, m, H-4 '), 3.88 (3H, s, OCH
3), 3.64 (2H, m, H-5 '), 3.57 (2H, m, H-α), 1.54 (3H, s, CH
3), 1.33 (3H, s, CH
3);
13C NMR (500MHz, DMSO-d
6): δ 173.24 (COOMe), 163.70 (C-2), 150.43 (C-4), 140.76 (C-6), 117.02,116.96 (＞CMe
2), 101.76 (C-1 '), 87.74 (C-5), 84.83 (C-2 '), 73.54 (C-3 '), 69.80 (C-4 '), 60.87 (C-5 '), 54.78 (OCH
3), 45.86 (C-α), 26.40 (CH
3), 26.86 (CH
3); ESI-MS (pos.): m/z 476 (M+H)
+ESI-MS (neg.): m/z 474 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
Like virus=Human immunodeficiency virus now
MT-4＝Human?leukenia?T?cell
CEM＝Human?lymphoblastoid?T?cell
ED
50=antiviral activity index
CD
50=cytotoxicity index
ED
50?CEM-TK- 5×10
-3M (CD
50?7×10
-6M)
CEM-SS 4×10
-3M (CD
60?8×10
-6M)
MT4 2×10
-3M (CD
60?8×10
-6M)
Embodiment 2: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-thiophosphoryl alanine methyl ester compound, wherein R is CH
3, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 62.4%.
Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 57.86,57.03;
1H NMR (500MHz, DMSO-d
6): δ 9.44,9.40 (bs, 1H, NH), 8.89 (1H, s, H-2), 8.67 (1H, s, H-8), 7.67 (2H, s, NH
2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH
3), 3.85 (2H, m, H-5 '), 3.67 (2H, m, H-α), 1.61 (3H, s, CH
3), 1.42 (3H, s, CH
3), 1.31,1.30 (3H, d,
3J=6, β-CH
3);
13C NMR (500MHz, DMSO-d
6): δ 180.24 (COOMe), 170.70 (C-2), 156.43 (C-4), 149.76 (C-6), 117.02,116.96 (＞CMe
2), 108.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH
3), 50.73 (C-α), 48.66 (C-β), 26.40 (CH
3), 26.86 (CH
3); ESI-MS (pos.): m/z 487 (M+H)
+ESI-MS (neg.): m/z 489 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 4×10
-3M (CD
50?2×10
-6M)
CEM-SS 6×10
-3M (CD
50?9×10
-6M)
MT4 8×10
-3M (CD
50?6×10
-6M)
Embodiment 3: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2, B is a VITAMIN B4.
The structural formula of compound:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.22g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 58.7%.
Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 58.64,58.33;
1H NMR (500MHz, DMSO-d
6): δ 9.21,9.15 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.88 (2H, s, NH
2), 7.15-7.35 (5H, m, Ph), 6.59 (1H, m, H-1 '), 4,55 (2H, m, H-2 ', 3 '), 4.37 (1H, m, H-4 '), 3.79 (3H, s, OCH
3), 3.66 (2H, m, H-5 '), 3.48 (2H, m, H-α), 2.32 (2H, m, H-β), 1.61 (3H, s, CH
3), 1.49 (3H, s, CH
3);
13C NMR (500MHz, DMSO-d
6): δ 174.24 (COOMe), 170.60 (C-2), 159.43 (C-4), 148.36 (Ph-jpso), 117.02,116.96 (＞CMe
2), 136.11 (C-6), 135.12 (Ph-para), 130.42 (Ph-ortho), 119.91 (Ph-meta), (108.23 C-1 '), 89.65 (C-5), 87.87 (C-2 '), 76.23 (C-3 '), (72.66 C-4 '), 61.23 (C-5 '), 56.82 (C-β), 56.78 (OCH
3), 52.73 (C-α), 26.40 (CH
3), 23.80 (CH
3); ESI-MS (pos.): m/z 565 (M+H)
+ESI-MS (neg.): m/z 563 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 8×10
-3M (CD
50?4×10
-6M)
CEM-SS 9×10
-3M (CD
50?7×10
-6M)
MT4 6×10
-3M (CD
50?8×10
-6M)
Embodiment 4: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CHCH
2, B is a VITAMIN B4.The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 55.2%.
Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 56.32,56.14;
1H NMR (500MHz, DMSO-d
6): δ 9.44,9.40 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.54 (2H, s, NH
2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH
3), 3.85 (2H, m, H-5 '), 3.67 (1H, m, H-α), 3.45 (1H, m, H-β), 1.61 (3H, s, CH
3), 1.42 (3H, s, CH
3), 1.31 (3H, d,
3J=6, CH
3), 1.25 (3H, d,
3J=6, CH
3);
13C NMR (500MHz, DMSO-d
6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), 117.02,116.96 (＞CMe
2), 108.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH
3), 50.73 (C-α), 48.66 (C-β), 26.40 (CH
3), 26.86 (CH
3); ESI-MS (pos.): m/z 518 (M+H)
+ESI-MS (neg.): m/z 516 (M-H)
-. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-3M (CD
50?2×10
-6M)
CEM-SS 6×10
-3M (CD
50?5×10
-6M)
MT4 8×10
-3M (CD
50?6×10
-6M)
Embodiment 5: preparation 2 ', 3 '-O-isopropylidene adenosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2, B is a VITAMIN B4.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene adenosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) 2 ', 3 '-O-isopropylidene adenosine synthetic with above-mentioned embodiment (1).
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene adenosine 5 '-thiophosphoryl leucine methyl esters, productive rate is 53.2%.
Spectral data is as follows:
31P NMR (DMSO-d
6, δ: ppm, J:Hz): δ 54.32,54.14;
1H NMR (500MHz, DMSO-d
6): δ 9.31,9.27 (bs, 1H, NH), 8.67 (1H, s, H-2), 8.58 (1H, s, H-8), 7.54 (2H, s, NH
2), 6.59 (1H, m, H-1 '), 4,78 (2H, m, H-2 ', 3 '), 4.22 (1H, m, H-4 '), 3.98 (3H, s, OCH
3), 3.85 (2H, m, H-5 '), 3.67 (1H, m, H-α), 3.45 (1H, m, H-β), 3.21 (1H, m, H-γ), 1.61 (3H, s, CH
3), 1.42 (3H, s, CH
3), 1.31 (3H, d,
3J=6, CH
3), 1.25 (3H, d,
3J=6, CH
3);
13C NMR (500MHz, DMSO-d
6): δ 172.24 (COOMe), 165.70 (C-2), 159.43 (C-4), 145.76 (C-6), 116.02,115.96 (＞CMe
2), 109.23 (C-1 '), 89.66 (C-5), 85.87 (C-2 '), 76.23 (C-3 '), 69.66 (C-4 '), 61.23 (C-5 '), 54.78 (OCH
3), 50.73 (C-α), 48.66 (C-β), 46.35 (C-γ), 23.40 (CH
3), 22.86 (CH
3); ESI-MS (pos.): m/z 531 (M+H)
+ESI-MS (neg.): m/z 529 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-3M (CD
50?2×10
-6M)
CEM-SS 2×10
-3M (CD
50?5×10
-6M)
MT4 5×10
-3M (CD
50?9×10
-6M)
Embodiment 6: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-thiophosphoryl glycine methyl ester compound, wherein R is H, B is a guanine.The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene guanosine is standby
Getting 1mol (286g) guanosine under the room temperature is suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system becomes clarification after about 1 hour, neutralize with 100ml strong aqua and 5.9L water, this moment product 2 ', 3 '-O-isopropylidene guanosine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, obtain after the drying product 2 ', 3 '-O-isopropylidene guanosine 240g (74%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.323g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) dichloro thiophosphoryl glycine methyl ester, productive rate is 58.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 56.68,56.39;
1H NMR (500MHz, D
2O): δ 8.18 (1H, s, H-8), 5.82,5.81 (1H, d,
3J=6.0, H-1 '), 4,89 (1H, m, H-2 '), 4.23 (1H, m, H-3 '), 3.23 (1H, m, H-4 '), 3.54 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 3.54 (2H, m, H-α), 1.66,1.65 (3H, d,
3J=6.0, β-CH
3), 1.35 (3H, s, CH
3), 1.33 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 172.31 (COOMe), 165.81 (C-2), 158.67 (C-6), 152.34 (C-4), 139.66 (C-8), 121.11 (＞CMe
2), 118.71 (C-5), 89.42 (C-4 '), 86.25 (C-1 '), 73.55 (C-3 '), 70.82 (C-2 '), 61.45 (C-5 '), 54.62 (OCH
3), 50.62 (C-β), 45.89 (C-α), 28.66 (CH
3), 26.1 5 (CH
3); ESI-MS (pos.): m/z 491 (M+H)
+ESI-MS (neg.): m/z 489 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 8×10
-3M (CD
50?6×10
-6M)
CEM-SS 3×10
-3M (CD
50?4×10
-6M)
MT4 6×10
-3M (CD
50?3×10
-6M)
Embodiment 7:2 ', 3 '-O-isopropylidene guanosine 5 '-preparation of (isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH
3, B is a guanine.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.140g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
4) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 52.3%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 55.88,55.37;
1H NMR (500MHz, D
2O): δ 8.07 (1H, s, H-8), 5.82,5.81 (1H, d,
3J=6.0, H-1 '), 4,51 (1H, m, H-2 '), 4.18 (1H, m, H-3 '), 3.94 (1H, m, H-4 '), 3.86 (3H, s, OCH
3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 1.35 (3H, s, CH
3), 1.33 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 178.31 (COOMe), 156.81 (C-2), 153.67 (C-6), 151.34 (C-4), 135.66 (C-8), 120.11 (＞CMe
2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 23.66 (CH
3), 23.15 (CH
3); ESI-MS (pos.): m/z 506 (M+H)
+ESI-MS (neg.): m/z 504 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-3M (CD
50?7×10
-5M)
CEM-SS 6×10
-3M (CD
50?9×10
-6M)
MT4 2×10
-4M (CD
50?7×10
-5M)
Embodiment 8: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2, B is a guanine.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.21g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 50.3%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.18,56.89;
1H NMR (500MHz, D
2O): δ 8.07 (1H, s, H-8), 7.25-7.41 (5H, m, Ph), 5.79,5.78 (1H, d,
3J=6.0, H-1 '), 4,41 (1H, m, H-2 '), 4.18 (1H, m, H-3 '), 3.94 (1H, m, H-4 '), 3.86 (3H, s, OCH
3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 2.27 (2H, m, H-β), 1.21 (3H, s, CH
3), 1.17 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 180.31 (COOMe), 166.81 (C-2), 161.67 (C-6), 158.34 (C-4), 151.26 (Ph-jpso), 135.66 (C-8), 130.07 (Ph-para), 128.23 (Ph-ortho), 118.45 (Ph-meta), 108.11 (＞CMe
2), 96.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 57.22 (C-β), 54.62 (OCH
3), 45.89 (C-α), 23.66 (CH
3), 23.15 (CH
3); ESI-MS (pos.): m/z 581 (M+H)
+ESI-MS (neg.): m/z 579 (M-H)
-. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-3M (CD
50?7×10
-5M)
CEM-SS 6×10
-3M (CD
50?9×10
-6M)
MT4 2×10
-4M (CD
50?7×10
-5M)
Embodiment 9: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH
2, B is a guanine.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.170g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 56.2%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 58.86,58.27;
1H NMR (500MHz, D
2O): δ 9.07 (1H, s, H-8), 6.82,6.81 (1H, d,
3J=6.0, H-1 '), 5,48 (1H, m, H-2 '), 4.95 (1H, m, H-3 '), 4.36 (1H, m, H-4 '), 3.86 (3H, s, OCH
3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 1.35 (3H, s, CH
3), 1.31 (3H, s, CH
3), 1.16,1.15 (3H, d,
3J=6.0, CH
3), 1.06,1.05 (3H, d,
3J=6.0, CH
3);
13C NMR (500MHz, D
2O): δ 178.31 (COOMe), 156.81 (C-2), 153.67 (C-6), 151.34 (C-4), 135.66 (C-8), 120.11 (＞CMe
2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 44.24 (C-β), 23.66 (CH
3), 23.15 (CH
3), 2 1.26 (CH
3), 20.05 (CH
3); ESI-MS (pos.): m/z 533 (M+H)
+ESI-MS (neg.): m/z 531 (M-H)
-. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-3M (CD
50?6×10
-5M)
CEM-SS 7×10
-2M (CD
50?9×10
-5M)
MT 44×10
-4M (CD
50?8×10
-5M)
Embodiment 10: preparation 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2, B is a guanine.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.180g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene guanosine synthetic with above-mentioned embodiment (6).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.31g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene guanosine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene guanosine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 50.2%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 55.86,55.27;
1H NMR (500MHz, D
2O): δ 9.07 (1H, s, H-8), 6.82,6.81 (1H, d,
3J=6.0, H-1 '), 5,48 (1H, m, H-2 '), 4.95 (1H, m, H-3 '), 4.36 (1H, m, H-4 '), 3.86 (3H, s, OCH
3), 3.68 (2H, m, H-5 '), 3.57 (2H, m, H-α), 3.45 (1H, m, H-β), 3.22 (1H, m, H-γ), 1.35 (3H, s, CH
3), 1.31 (3H, s, CH
3), 1.16,1.15 (3H, d,
3J=6.0, CH
3), 1.06,1.05 (3H, d,
3J=6.0, CH
3);
13C NMR (500MHz, D
2O): δ 178.31 (COOMe), 166.81 (C-2), 159.67 (C-6), 156.34 (C-4), 134.66 (C-8), 17.11 (＞CMe
2), 116.71 (C-5), 86.42 (C-4 '), 85.25 (C-1 '), 73.74 (C-3 '), 70.42 (C-2 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 44.24 (C-β), 41.24 (C-γ), 21.26 (CH
3), 20.05 (CH
3); ESI-MS (pos.): m/z 548 (M+H)
+ESI-MS (neg.): m/z 546 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-3M (CD
50?6×10
-5M)
CEM-SS 8×10
-3M (CD
50?8×10
-6M)
MT4 8×10
-4M (CD
50?2×10
-5M)
Embodiment 11: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene cytidine is standby
Get 1mol (244g) guanosine under the room temperature and be suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system change clarification after about 1 hour, adding 1mol (84g) NaHCO
3Neutralize, continue to stir 15 minutes, filter the back solids washed with acetone, with obtain behind filtrate and the washings evaporate to dryness product 2 ', 3 '-O-isopropylidene cytidine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, obtain after the drying product 2 ', 3 '-O-isopropylidene cytidine 263g (92%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 60.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.68,56.26;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.1 5 (1H, H-2 '), 1.1 5 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 451 (M+H)
+ESI-MS (neg.): m/z 449 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 8×10
-4M (CD
50?6×10
-5M)
CEM-SS 6×10
-3M (CD
50?4×10
-5M)
MT4 6×10
-4M (CD
50?3×10
-6M)
Embodiment 12: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH
3, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and is cooled to-4～-8 ℃, with (isopropoxy replacement) dichloro of 1mmol (0.17g)
Sulphur phosphorus is dissolved in the dry tetrahydrofuran (THF) of crossing (THF), is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 62.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 55.66,55.31;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.88 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.68 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 1.45 (3H, d,
3J=6, H-β), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 50.86 (C-β), 45.89 (C-α), 39.82 (C-2 '), 27.66 (CH
3), 24.15 (CH
3); ESI-MS (pos.): m/z 464 (M+H)
+ESI-MS (neg.): m/z 462 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-3M (CD
50?5×10
-5M)
CEM-SS 5×10
-3M (CD
50?8×10
-6M)
MT4 7×10
-4M (CD
50?3×10
-6M)
Embodiment 13: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) dichloro thiophosphoryl phenylalanine methyl ester compound, wherein R is C
6H
5CH
2, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.22g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 56.7%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 58.71,58.13;
1H NMR (500MHz, D
2O): δ 8.22,8.21 (1H, d,
3J=5.5, H-6), 7.29-7.41 (5H, m, Ph), 7.03 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.66 (1H, m, H-3 '), 3.82 (1H, m, H-4 '), 3.58 (3H, s, OCH
3), 3.44 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 2.27 (1H, m, H-β), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 180.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 146.36 (Ph-jpso), 117.02,136.11 (C-6), 137.12 (Ph-para), 136.42 (Ph-ortho), 122.91 (Ph-meta), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 50.86 (C-β), 45.89 (C-α), 39.82 (C-2 '), 27.66 (CH
3), 24.15 (CH
3); ESI-MS (pos.): m/z 540 (M+H)
+ESI-MS (neg.): m/z 538 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-4M (CD
50?8×10
-5M)
CEM-SS 6×10
-4M (CD
50?8×10
-5M)
MT4 8×10
-4M (CD
50?7×10
-6M)
Embodiment 14: preparation 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH
2, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 52.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 56.98,56.21;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.63 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.61 (1H, m, H-3 '), 3.69 (1H, m, H-4 '), 3.76 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.89 (2H, m, H-α), 2.66 (1H, H-2 '), 2.14 (1H, m, H-β), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 50.86 (C-β), 49.89 (C-α), 46.21 (C-β), 39.82 (C-2 '), 24.37 (CH
3), 23.22 (CH
3); ESI-MS (pos.): m/z 506 (M+H)
+ESI-MS (neg.): m/z 504 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-4M (CD
50?5×10
-7M)
CEM-SS 5×10
-4M (CD
50?6×10
-6M)
MT4 8×10
-3M (CD
50?3×10
-5M)
Embodiment 15.2 ', 3 '-O-isopropylidene cytidine 5 '-preparation of (isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2, B is a cytosine(Cyt).
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene cytidine synthetic with above-mentioned embodiment 11.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, with 1mmol (0.282g) be dissolved in 2 in the pyridine ', 3 '-O-isopropylidene cytidine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene cytidine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 55.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 54.36,53.56;
1H NMR (500MHz, D
2O): δ 7.96,7.95 (1H, d,
3J=6, H-6), 6.36 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.78 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.76 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 3.22 (2H, m, H-α), 2.96 (1H, H-2 '), 2.77 (1H, m, H-β), 2.39 (1H, m, H-γ), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 185.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 145.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 88.25 (C-1 '), 85.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.88 (OCH
3), 50.86 (C-β), 48.89 (C-α), 46.21 (C-γ), 39.82 (C-2 '), 23.39 (CH
3), 23.01 (CH
3); ESI-MS (pos.): m/z 508 (M+H)
+ESI-MS (neg.): m/z 506 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 5×10
-3M (CD
50?6×10
-6M)
CEM-SS 8×10
-4M (CD
50?9×10
-6M)
MT4 7×10
-4M (CD
50?8×10
-6M)
Embodiment 16: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester compound, wherein R is H, B is a uridylic.The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the glycine methyl ester hydrochloride of 1mmol (0.125g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) preparation 2 ', 3 '-O-isopropylidene uridine is standby
Get 1mol (244g) uridine under the room temperature and be suspended in the acetone of 2L, stir down to wherein adding 1.1mol (210g) tosic acid and 4mol (640ml) triethyl orthoformate, system change clarification after about 1 hour, adding 1mol (84g) NaHCO
3Neutralize, continue to stir 15 minutes, filter the back solids washed with acetone, with obtain behind filtrate and the washings evaporate to dryness product 2 ', 3 '-O-isopropylidene uridine slowly separates out, standing over night under the room temperature, after the filtration again with ice-water mixture thorough washing, filter rear filtrate through underpressure distillation obtain product 2 ', 3 '-O-isopropylidene uridine 256g (90%).
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl glycine methyl ester, productive rate is 63.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 57.68,56.26;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 480 (M+H)
+ESI-MS (neg.): m/z 478 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 8×10
-3M (CD
50?9×10
-5M)
CEM-SS 9×10
-3M (CD
50?8×10
-5M)
MT4 8×10
-4M (CD
50?5×10
-6M)
Embodiment 17: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is CH
3, B is a uridylic.
The structural formula of compound is:
The synthesis step of compound:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the alanine methyl ester hydrochloride of 1mmol (0.14g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl alanine methyl ester, productive rate is 58.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 58.32,57.68;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 466 (M+H)
+ESI-MS (neg.): m/z 464 (M-H)
-. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 8×10
-4M (CD
50?9×10
-4M)
CEM-SS 7×10
-4M (CD
50?6×10
-5M)
MT4 8×10
-3M (CD
50?5×10
-5M)
Embodiment 18.2 ', 3 '-O-isopropylidene uridine 5 '-preparation of (isopropoxy replacement) thiophosphoryl alanine methyl ester compound, wherein R is C
6H
5CH
2, B is a uridylic.The structural formula of compound:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the phenylalanine methyl ester hydrochloride of 1mmol (0.21g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl phenylalanine methyl ester, productive rate is 51.4%.Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 60.32,59.68;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 542 (M+H)
+ESI-MS (neg.): m/z 540 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-4M (CD
50?7×10
-5M)
CEM-SS 9×10
-3M (CD
50?9×10
-5M)
MT4 5×10
-3M (CD
50?6×10
-4M)
Embodiment 19: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester compound, wherein R is (CH
3)
2CH
2, B is a uridylic.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the valine methyl ester hydrochloride of 1mmol (0.17g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O-isopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, 1mmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl valine methyl ester, productive rate is 66.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 60.82,59.98;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 542 (M+H)
+ESI-MS (neg.): m/z 540 (M-H)
-. the anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-4M (CD
50?7×10
-5M)
CEM-SS 9×10
-3M (CD
50?9×10
-5M)
MT4 5×10
-3M (CD
50?6×10
-4M)
Embodiment 20: preparation 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl compound, wherein R is (CH
3)
2CHCH
2, B is a uridylic.
The structural formula of compound is:
The synthesis step of compound is as follows:
1) under the nitrogen protection, cryosel is bathed and to be cooled to-4～-8 ℃, and (isopropoxy replacements) two compd 22/190s of 1mmol (0.17g) are dissolved in the tetrahydrofuran (THF) (THF) of dry mistake, is mixed with the solution of 1mol/L.
2) in above-mentioned solution, add the leucine methyl ester hydrochloride of 1mmol (0.18g), slowly drip 2mmol (0.2g) triethylamine after stirring.
3) 2 ', 3 '-O diisopropylidene uridine synthetic with above-mentioned experiment 16.
4) follow the tracks of above-mentioned second reaction process that goes on foot with nuclear magnetic resonance analyser (NMR), after treating that (isopropoxy replacement) two compd 22/190 total overall reactions finish, lmmol (0.284g) has been dissolved in 2 among the THF ', 3 '-O-isopropylidene uridine slowly splashes in the above-mentioned system, and 1mmol (0.1g) triethylamine is continued to drip in the back that stirs.
5) finish with the NMR monitoring reaction after, filter, rotary distillation removes and desolvates and other low-boiling point materials, is hydrolyzed with ammoniacal liquor at last.After hydrolysis is finished, carry out column chromatography for separation with silicagel column, eluent is a Virahol: ammoniacal liquor: water=33: 1: 1 can obtain product 2 ', 3 '-O-isopropylidene uridine 5 '-(isopropoxy replacement) thiophosphoryl leucine methyl esters, productive rate is 62.4%.
Spectral data is as follows:
31P NMR (D
2O, δ: ppm, J:Hz): δ 59.87,59.68;
1H NMR (500MHz, D
2O): δ 7.90,7.89 (1H, d,
3J=5.5, H-6), 6.24 (1H, m, H-1 '), 5.84,5.83 (1H, d,
3J=6, H-5), 4.28 (1H, m, H-3 '), 3.88 (1H, m, H-4 '), 3.71 (3H, s, OCH
3), 3.65 (2H, m, H-5 '), 2.69 (2H, m, H-α), 2.15 (1H, H-2 '), 1.15 (3H, s, CH
3), 1.07 (3H, s, CH
3);
13C NMR (500MHz, D
2O): δ 177.63 (COOMe), 165.82 (C-4), 158.13 (C-2), 140.34 (C-6), 121.11 (＞CMe
2), 93.71 (C-5), 87.25 (C-1 '), 84.42 (C-4 '), 70.42 (C-3 '), 61.45 (C-5 '), 54.62 (OCH
3), 45.89 (C-α), 39.82 (C-2 '), 28.66 (CH
3), 26.15 (CH
3); ESI-MS (pos.): m/z 542 (M+H)
+ESI-MS (neg.): m/z 540 (M-H)
-.
The anti-love of this compound in cem cell and MT-4 cell is virus-1 activity experiment now
ED
50?CEM-TK- 6×10
-4M (CD
50?7×10
-5M)
CEM-SS 9×10
-3M (CD
50?9×10
-5M)
MT4 5×10
-3M (CD
50?6×10
-4M)
Claims (2)
1, a kind of 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound, it is characterized in that the structural formula of this compound is:
R is H, CH in the said structure formula
3, C
6H
5CH
2, (CH
3)
2CH
2Or (CH
3)
2CHCH
2In any; B is any in VITAMIN B4, guanine, cytosine(Cyt) or the uridylic.
2, a kind of preparation method of compound as claimed in claim 1 is characterized in that this method comprises the steps:
1) under nitrogen protection, cryosel is bathed under the condition that is cooled to-4～-8 ℃, and raw material (isopropoxy replacement) two compd 22/190s are dissolved in the dry tetrahydrofuran (THF) of crossing, and is mixed with the solution that concentration is 0.8～1.0mol/L;
2) add amino acid methyl ester hydrochloride with the amount of raw material same substance in above-mentioned solution, slowly drip acid binding agent second triamine after stirring, the add-on of acid binding agent is 2 times of amount of raw material;
3) preparation 2 ', 3 '-O-isopropylidene nucleosides solution for standby:
Get nucleosides under the room temperature and be suspended in the solution that is made into 0.5～1.0mol/L in the exsiccant solvent, stir down to wherein adding and the tosic acid of amount of substances such as nucleosides and the triethyl orthoformate of four times of amount of substances, after system becomes clarification, neutralize with big water gaging with strong aqua and to make PH=7～8, then above-mentioned solution decompression is concentrated, in refrigerator standing over night can obtain product 2 ', 3 '-O-isopropylidene nucleosides.With make 2 ', 3 '-be dissolved in the solution that is mixed with 0.5～1.0 mol/L in the solvent after the O-isopropylidene nucleosides suction filtration drying;
4) follow the tracks of above-mentioned second reaction process that goes on foot, after treating that the raw material total overall reaction finishes, will with 2 of above-mentioned the 3rd step preparation of the amount of raw material same substance ', 3 '-O-isopropylidene nucleosides solution slowly splashes in the solution in second step, and the back that stirs continue to drip the acid binding agent with the amount of raw material same substance;
5) after monitoring reaction is finished, filter, rotary distillation removes and desolvates and other low-boiling point materials, be hydrolyzed with ammoniacal liquor at last, recycle silicon glue post carries out column chromatography for separation, can obtain 2 ', 3 '-O-isopropylidene nucleosides 5 '-(isopropoxy replacement) thiophosphoryl amino acid ester compound.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011307846A CN1159332C (en) | 2001-08-24 | 2001-08-24 | Thiophosphoryl amino acid ester compound and its prepn process |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011307846A CN1159332C (en) | 2001-08-24 | 2001-08-24 | Thiophosphoryl amino acid ester compound and its prepn process |
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| Publication Number | Publication Date |
|---|---|
| CN1337401A true CN1337401A (en) | 2002-02-27 |
| CN1159332C CN1159332C (en) | 2004-07-28 |
Family
ID=4670143
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|---|---|---|---|
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8871737B2 (en) | 2010-09-22 | 2014-10-28 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US8916538B2 (en) | 2012-03-21 | 2014-12-23 | Vertex Pharmaceuticals Incorporated | Solid forms of a thiophosphoramidate nucleotide prodrug |
| US8980865B2 (en) | 2011-12-22 | 2015-03-17 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US9012427B2 (en) | 2012-03-22 | 2015-04-21 | Alios Biopharma, Inc. | Pharmaceutical combinations comprising a thionucleotide analog |
-
2001
- 2001-08-24 CN CNB011307846A patent/CN1159332C/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8871737B2 (en) | 2010-09-22 | 2014-10-28 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US9278990B2 (en) | 2010-09-22 | 2016-03-08 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US8980865B2 (en) | 2011-12-22 | 2015-03-17 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US9605018B2 (en) | 2011-12-22 | 2017-03-28 | Alios Biopharma, Inc. | Substituted nucleotide analogs |
| US8916538B2 (en) | 2012-03-21 | 2014-12-23 | Vertex Pharmaceuticals Incorporated | Solid forms of a thiophosphoramidate nucleotide prodrug |
| US9394330B2 (en) | 2012-03-21 | 2016-07-19 | Alios Biopharma, Inc. | Solid forms of a thiophosphoramidate nucleotide prodrug |
| US9856284B2 (en) | 2012-03-21 | 2018-01-02 | Alios Biopharma, Inc. | Solid forms of a thiophosphoramidate nucleotide prodrug |
| US9012427B2 (en) | 2012-03-22 | 2015-04-21 | Alios Biopharma, Inc. | Pharmaceutical combinations comprising a thionucleotide analog |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1159332C (en) | 2004-07-28 |
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